fishes Article Trypanorhynch Assemblages Indicate Ecological and Phylogenetical Attributes of Their Elasmobranch Final Hosts Harry W. Palm 1,*, Irfan Yulianto 1,2 and Uwe Piatkowski 3 1 Aquaculture and Sea-Ranching, Agricultural and Environmental Sciences, University of Rostock, Justus-von-Liebig Weg 6, 18059 Rostock, Germany 2 Marine Fisheries Department, Faculty of Fisheries and Marine Sciences, Bogor Agricultural University (IPB), Jl. Raya Darmaga Kampus IPB Darmaga, 16680 Bogor, Indonesia; [email protected] 3 GEOMAR, Helmholtz Centre for Ocean Research Kiel, Düsternbrooker Weg 20, 24105 Kiel, Germany; [email protected] * Correspondence: [email protected]; Tel.: +49-381-498-3730 Academic Editor: Maria Angeles Esteban Received: 7 February 2017; Accepted: 12 June 2017; Published: 17 June 2017 Abstract: This study explores non-metric multidimensional scaling (nMDS) as a tool for investigating parasites as indicators of the elasmobranch biology. An attractive feature of nMDS is its ability to allow assemblage-level parasite data to be simultaneously applied to questions of host biology. This method was examined using the tapeworm order Trypanorhyncha Diesing, 1863, which is known to be transmitted among their hosts through the marine food web (via predation), can unambiguously be identified in the intermediate and final hosts, and has the potential as an indicator of the host feeding biology. Our analyses focused on trypanorhynch assemblages in elasmobranchs as definitive hosts. The relationships between trypanorhynch assemblages and the depth, feeding ecology, habitat, and phylogeny for all sharks were complex, but we found that depth distribution, diet composition and habitat type were the major influencing factors. Several species of sharks showed different characters than known from their descriptions that could be attributed to the change of shark behavior or the trypanorhynch host path. The relationship between the trypanorhynch assemblage and factors for carcharhiniform species alone was more robust than for all sharks. In the carcharhiniform analysis, the relationship between habitat type and trypanorhynch assemblage was most remarkable. Overlapping host ecology was evident even in phylogenetically-distant related hosts. Keywords: biological indicator; cestodes; classification; elasmobranchs; host specificity; host switch; phylogeny; feeding ecology; Trypanorhyncha 1. Introduction Elasmobranchs are difficult to observe in nature and information on rare or less-frequently caught species is scarce. This especially concerns aspects of their ecology, main habitat, depth range, and most important prey items. Their prey is rapidly digested and affiliation, e.g., to mainly fish or invertebrate-feeding activities, is difficult. Fish parasites have been widely used as biological indicators for the host ecology, including their zoogeographical distribution, migration, and feeding behavior [1]. However, the methods applied for the elasmobranchs are, so far, limited due to often restricted and unpredictable catches and less availability of specimens to study. The cestode order Trypanorhyncha Diesing, 1863, is considered to be a basal tapeworm group of elasmobranchs. Its members can be characterized by a scolex with a tentacular apparatus that Fishes 2017, 2, 8; doi:10.3390/fishes2020008 www.mdpi.com/journal/fishes Fishes 2017, 2, 8 2 of 16 can be used for classification [2–5]. The most recent revision of the trypanorhynchs subdivided the order into five different superfamilies and 15 families [6]. The first large-scale molecular phylogenetic analyses of this group included trypanorhynch species belonging to all superfamilies and 12 of the 15 recognized families [7,8]. The analysis of complete simple sequence repeats DNA (ssrDNA) and partial long sequence repeat DNA (lsrDNA) (D1–D3) revealed high resolution and the presence of a basal split, representing the shark- and ray-inhabiting lineages as new suborders. These results gave, in part, support for the trypanorhynch classification by Palm [6] as well as more insight into the intrafamily phylogenetic relationships within the Trypanorhyncha. The host-parasite-checklist of these cestodes indicated that some of the superfamilies and families seem to have host preferences [6], while the patterns of others can be explained by host switches, even between rajiform and selachiform elasmobranchs [7]. The ~300 described species of the order Trypanorhyncha so far exhibit the following general life-cycle pattern [6,9–11]. The first intermediate host becomes infected when it consumes an oncosphere or free-swimming coracidium larva. The first parasitic larval stage is a procercoid; the limited available data suggest that this stage parasitizes copepods. Second intermediate hosts include a wide array of marine animals, but invertebrates and teleosts figure particularly prominent at this level. Unlike most groups of tapeworms, the trypanorhynch final larval stage, though having a different morphology (plerocercoid, plerocercus, merocercoid), is generally identifiable to species because it exhibits the adult hook pattern, a pattern on which much of the taxonomy of the group is based [4–6]. Some trypanorhynchs use a paratenic host following the final intermediate host; this host serves to bridge food-web gaps. As adults, trypanorhynchs parasitize the spiral intestine of sharks and rays (elasmobranchs). Fortunately, it is common for individual elasmobranch species to host multiple trypanorhynch species and, thus, assemblage-level data are readily available for many elasmobranchs that serve as definitive hosts. Our study uses non-metric multidimensional scaling (nMDS) as a tool for investigating parasites as indicators for selected ecological attributes and the phylogeny of their elasmobranch final hosts. A particularly novel and attractive feature of nMDS is the possibility to allow assemblage-level parasite data to be simultaneously applied to questions of host biology. The method was examined using trypanorhynch tapeworm assemblages, which have obvious potential as indicators of host feeding biology in the elasmobranch. The complex life-cycles of these parasites, each of which involves a minimum of three different hosts and three distinct life-cycle stages that vary in their degree of host specificity [11] also suggest trypanorhynchs as appropriate indicators of other aspects of their host biology. Among elasmobranchs, sharks (Selachii) were specifically chosen because they vary substantially in diet and habitat, which allows a wide spectrum of aspects of host biology to be explored. Within the Selachii, more detailed analyses were conducted on sharks of the order Carcharhiniformes (ground sharks). Comprehensive trypanorhynch data were available for this shark order, and provided a unique opportunity to perform nMDS analyses on a more precise taxonomic level. Factors concerning the shark biology included (1) taxonomy (order for the Selachii analyses, or family for the Carcharhiniform analyses); (2) habitat (benthic vs. pelagic); (3) depth distribution (above or below 200 m depth); and (4) diet (primarily invertebrates vs. primarily teleosts). Trypanorhynch assemblage dependence on these four factors was assessed by superimposing the different variables into the two-dimensional ordination plots resulting from nMDS analysis of the hierarchical agglomerative classification of shark species generated from their trypanorhynch assemblages. Trypanorhynch assemblages were considered to be appropriate as indicators of the biological factors that were found to be significantly correlated with the agglomerative classification. Fishes 2017, 2, 8 3 of 16 Fishes 2017, 2, 8 3 of 18 2.2. ResultsResults 2.1.2.1. AllAll SharksSharks TheThe resultresult ofof thethe hierarchical,hierarchical, agglomerative classification classification (CLUSTER) (CLUSTER) of of trypanorhynch trypanorhynch assemblagesassemblages ofof 50 species of Selachii revealed revealed five five major major clusters clusters of of shark shark species species at at an an arbitrary arbitrary dissimilaritydissimilarity level of 78% (Figure 11).). TheThe two-dimensionaltwo-dimensional ordination plots plots resulti resultingng from from nMDS nMDS of of thisthis agglomerativeagglomerative classification,classification, with with sharks sharks coded coded for for taxonomic taxonomic order, order, diet, diet, habitat, habitat, and and depth depth distributiondistribution areare shownshown in FigureFigure2 .2. TheThe analysisanalysis ofof similaritiessimilarities (ANOSIM)(ANOSIM) revealed significant significant differencesdifferences inin trypanorhynchtrypanorhynch assemblage composition composition between between sharks sharks that that primarily primarily consume consume invertebratesinvertebrates versusversus thosethose that primarily consume teleosts teleosts (global (global RR == 0.279; 0.279; pp << 0.01), 0.01), between between those those fromfrom benthicbenthic andand pelagicpelagic habitats (global R = 0.275; 0.275; pp << 0.01), 0.01), and and between between sharks sharks oc occurringcurring in in deep deep versusversus shallowshallow water water (global (globalR =R 0.327;= 0.327;p < p 0.01), < 0.01), thus thus supporting supporting the significance the significance of the of major the clustersmajor resultingclusters resulting from the from classification the classification and ordination and ordination methods. methods. With respect With torespect taxonomy, to taxonomy, the ANOSIM the showedANOSIM significant showed significant differences differences among